IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ http://www.ebi.ac.uk/ ger-2011-1 Adaptor protein Nck1 interacts with p120 Ras GTPase-activating protein and regulates its activity. human-hep_g2 Homo sapiens hepatocellular carcinoma (HCC) cells (p53-wt). hep_g2 human hepatocellular carcinoma (HCC) cells (p53-wt). anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Ger M., Zitkus Z., Valius M. RasGAP activity could not be captures as source of H-Ras could not be traced. marija.ger@bchi.vu.lt, zigmantas.zitkus@bchi.vu.lt, mindaugas.valius@bchi.vu.lt 2011-11-01: Requested by Mindaugas Valius, Proteomics Centre, Vilnius University Institute of Biochemistry, Mokslininku st. 12, Vilnius LT-08662, Lithuania Cell. Signal. (0898-6568) 2011 Only protein-protein interactions imex curation ger-2011-2 Adaptor protein Nck1 interacts with p120 Ras GTPase-activating protein and regulates its activity. human-hep_g2 Homo sapiens hepatocellular carcinoma (HCC) cells (p53-wt). hep_g2 human hepatocellular carcinoma (HCC) cells (p53-wt). anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Ger M., Zitkus Z., Valius M. RasGAP activity could not be captures as source of H-Ras could not be traced. marija.ger@bchi.vu.lt, zigmantas.zitkus@bchi.vu.lt, mindaugas.valius@bchi.vu.lt 2011-11-01: Requested by Mindaugas Valius, Proteomics Centre, Vilnius University Institute of Biochemistry, Mokslininku st. 12, Vilnius LT-08662, Lithuania Cell. Signal. (0898-6568) 2011 Only protein-protein interactions imex curation ger-2011-3 Adaptor protein Nck1 interacts with p120 Ras GTPase-activating protein and regulates its activity. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Ger M., Zitkus Z., Valius M. RasGAP activity could not be captures as source of H-Ras could not be traced. marija.ger@bchi.vu.lt, zigmantas.zitkus@bchi.vu.lt, mindaugas.valius@bchi.vu.lt 2011-11-01: Requested by Mindaugas Valius, Proteomics Centre, Vilnius University Institute of Biochemistry, Mokslininku st. 12, Vilnius LT-08662, Lithuania Cell. Signal. (0898-6568) 2011 Only protein-protein interactions imex curation ger-2011-4 Adaptor protein Nck1 interacts with p120 Ras GTPase-activating protein and regulates its activity. in vitro In vitro far western blotting far western blotting Affinity blotting Affinity blotting western blot western blot Immuno blot Immuno blot Ger M., Zitkus Z., Valius M. RasGAP activity could not be captures as source of H-Ras could not be traced. marija.ger@bchi.vu.lt, zigmantas.zitkus@bchi.vu.lt, mindaugas.valius@bchi.vu.lt 2011-11-01: Requested by Mindaugas Valius, Proteomics Centre, Vilnius University Institute of Biochemistry, Mokslininku st. 12, Vilnius LT-08662, Lithuania Cell. Signal. (0898-6568) 2011 Only protein-protein interactions imex curation ger-2011-5 Adaptor protein Nck1 interacts with p120 Ras GTPase-activating protein and regulates its activity. in vitro In vitro ms of complexes mass spectrometry studies of complexes ms of complexes ms participant Identification by mass spectrometry ms participant Ger M., Zitkus Z., Valius M. RasGAP activity could not be captures as source of H-Ras could not be traced. marija.ger@bchi.vu.lt, zigmantas.zitkus@bchi.vu.lt, mindaugas.valius@bchi.vu.lt 2011-11-01: Requested by Mindaugas Valius, Proteomics Centre, Vilnius University Institute of Biochemistry, Mokslininku st. 12, Vilnius LT-08662, Lithuania Cell. Signal. (0898-6568) 2011 Only protein-protein interactions imex curation nck1_human Cytoplasmic protein NCK1 NCK1 NCK NCK adaptor protein 1 SH2/SH3 adaptor protein NCK-alpha protein protein human Homo sapiens Human MAEEVVVVAKFDYVAQQEQELDIKKNERLWLLDDSKSWWRVRNSMNKTGFVPSNYVERKNSARKASIVKNLKDTLGIGKVKRKPSVPDSASPADDSFVDPGERLYDLNMPAYVKFNYMAEREDELSLIKGTKVIVMEKCSDGWWRGSYNGQVGWFPSNYVTEEGDSPLGDHVGSLSEKLAAVVNNLNTGQVLHVVQALYPFSSSNDEELNFEKGDVMDVIEKPENDPEWWKCRKINGMVGLVPKNYVTVMQNNPLTSGLEPSPPQCDYIRPSLTGKFAGNPWYYGKVTRHQAEMALNERGHEGDFLIRDSESSPNDFSVSLKAQGKNKHFKVQLKETVYCIGQRKFSTMEELVEHYKKAPIFTSEQGEKLYLVKHLS rasa1_human Ras GTPase-activating protein 1 Ras p21 protein activator p120GAP RASA1 RASA GAP protein protein human Homo sapiens Human MMAAEAGSEEGGPVTAGAGGGGAAAGSSAYPAVCRVKIPAALPVAAAPYPGLVETGVAGTLGGGAALGSEFLGAGSVAGALGGAGLTGGGTAAGVAGAAAGVAGAAVAGPSGDMALTKLPTSLLAETLGPGGGFPPLPPPPYLPPLGAGLGTVDEGDSLDGPEYEEEEVAIPLTAPPTNQWYHGKLDRTIAEERLRQAGKSGSYLIRESDRRPGSFVLSFLSQMNVVNHFRIIAMCGDYYIGGRRFSSLSDLIGYYSHVSCLLKGEKLLYPVAPPEPVEDRRRVRAILPYTKVPDTDEISFLKGDMFIVHNELEDGWMWVTNLRTDEQGLIVEDLVEEVGREEDPHEGKIWFHGKISKQEAYNLLMTVGQVCSFLVRPSDNTPGDYSLYFRTNENIQRFKICPTPNNQFMMGGRYYNSIGDIIDHYRKEQIVEGYYLKEPVPMQDQEQVLNDTVDGKEIYNTIRRKTKDAFYKNIVKKGYLLKKGKGKRWKNLYFILEGSDAQLIYFESEKRATKPKGLIDLSVCSVYVVHDSLFGRPNCFQIVVQHFSEEHYIFYFAGETPEQAEDWMKGLQAFCNLRKSSPGTSNKRLRQVSSLVLHIEEAHKLPVKHFTNPYCNIYLNSVQVAKTHAREGQNPVWSEEFVFDDLPPDINRFEITLSNKTKKSKDPDILFMRCQLSRLQKGHATDEWFLLSSHIPLKGIEPGSLRVRARYSMEKIMPEEEYSEFKELILQKELHVVYALSHVCGQDRTLLASILLRIFLHEKLESLLLCTLNDREISMEDEATTLFRATTLASTLMEQYMKATATQFVHHALKDSILKIMESKQSCELSPSKLEKNEDVNTNLTHLLNILSELVEKIFMASEILPPTLRYIYGCLQKSVQHKWPTNTTMRTRVVSGFVFLRLICPAILNPRMFNIISDSPSPIAARTLILVAKSVQNLANLVEFGAKEPYMEGVNPFIKSNKHRMIMFLDELGNVPELPDTTEHSRTDLSRDLAALHEICVAHSDELRTLSNERGAQQHVLKKLLAITELLQQKQNQYTKTNDVR rasa1-nck1 1189199 n/a 1189204 unspecified role unspecified role prey prey n/a 1189205 unspecified role unspecified role bait bait physical association physical association false false false Fig. 1B nck1-rasa1 1189199 n/a 1189205 unspecified role unspecified role prey prey n/a 1189204 unspecified role unspecified role bait bait physical association physical association false false false Fig. 1B nck1-rasa1-1 1189200 n/a 1189204 unspecified role unspecified role bait bait P16333:p.Trp38Lys mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P16333:p.[Trp38Lys;Trp143Lys;Trp229Lys;Arg308Lys] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false P16333:p.[Trp38Lys;Trp229Lys;Arg308Lys] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false P16333:p.Trp229Lys mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P16333:p.[Trp38_Trp39delinsLysLys;Trp143Lys;Trp229Lys] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false P16333:p.[Trp143Lys;Trp229Lys;Arg308Lys] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false n/a 1189205 unspecified role unspecified role prey prey physical association physical association false false false Fig. 2A, 2B nck1-rasa1-2 1189201 n/a 1189204 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false sh3 region sufficient to bind sufficient binding region sufficient to bind range range range range false ecoli Escherichia coli (strain K12) n/a 1189205 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false human-hep_g2 Homo sapiens hepatocellular carcinoma (HCC) cells (p53-wt). hep_g2 human hepatocellular carcinoma (HCC) cells (p53-wt). 1189227 1189225 direct interaction direct interaction false false false Fig. 2C nck1-rasa1-3 1189202 n/a 1189204 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false sh3 region sufficient to bind sufficient binding region sufficient to bind range range range range false ecoli Escherichia coli (strain K12) n/a 1189205 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false human-hep_g2 Homo sapiens hepatocellular carcinoma (HCC) cells (p53-wt). hep_g2 human hepatocellular carcinoma (HCC) cells (p53-wt). direct interaction direct interaction false false false Fig. 3A nck1-rasa1-4 1189203 n/a 1189204 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) n/a 1189205 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false human-hep_g2 Homo sapiens hepatocellular carcinoma (HCC) cells (p53-wt). hep_g2 human hepatocellular carcinoma (HCC) cells (p53-wt). direct interaction direct interaction false false false Fig. 3C Molecular weight of complex = 51 474 Da.